Objective To investigate clinical diagnosis，timing of surgery and perioperative therapeutic strategies for blood culture-negative infective endocarditis （IE）. Methods Clinical data of 240 IE patients who were admitted tWuhan Asia Heart Hospital between July 2008 and July 2012 were retrospectively analyzed. According to their blood cultureresults，all the patients were divided into blood culture-negative group and blood culture-positive group. In the blood culture-negative group，there were 158 patients including 88 male and 70 female patients with their age of 51.3±10.1 years. In the blood culture-positive group，there were 82 patients including 45 male and 37 female patients with their age of 48.9±9.8 years. All the patients underwent surgical treatment，and the surgical procedures included complete vegetations excision，debridement of infected valves，removal of necrotic tissue around the annulus，and concomitant heart valve replacement or intracardiac repair. Postoperatively，all the patients received routine monitoring in ICU，cardiac glycosides，diuretics，other symptomatic treatment and adequate dosages of antibiotics for 4-6 weeks. Results Four patients died postoperatively in this study including 1 patient for low cardiac output syndrome and 3 patients for multiple organ dysfunction syndrome，1 patient in the blood culture-positive group and 3 patients in the blood culture-negative group respectively. There was no statistical difference in surgical mortality between the 2 groups （χ2=0.15，P=0.70）. All the other patients were discharged successfully and followed up for 6 to 36 months with the median follow-up time of 22 months. During follow-up, 2 patients died including 1 patient for cerebral infarction 2 years after surgery and another patient for cerebral hemorrhage 3 yearsafter surgery. Conclusion Patients with blood culture-negative IE should receive adequate dosage and duration of broad-spectrum antibiotics to control the infection rapidly, and aggressive surgical therapy to decrease in-hospital mortality and improve their quality of life and prognosis.